JP5332613B2 - Manufacturing method of resin molding - Google Patents

Abstract

In order to prevent a protein or peptide from adsorbing on a resin molding, the resin molding is brought into contact with an aqueous solution of a nonionic surfactant, and then the resin molding is irradiated with radiation rays and also a concentration of the surfactant in the aqueous solution is made to be 0.05 times to 500 times a critical micelle concentration of the surfactant at 25°C.

Description

  The present invention relates to a method for producing a resin molding particularly suitable for protein and / or peptide treatment.

  In recent years, proteomic analysis research (proteomics) has begun to attract attention as post-genomic research. Since protein, which is a gene product, is thought to be directly linked to disease pathology rather than gene, research results of proteome analysis that comprehensively examines proteins are expected to be widely applicable to diagnosis and treatment.

  The reason why proteome analysis has made rapid progress is technically that high-speed structural analysis by mass spectrometer (MS) has become possible. Practical applications such as MALDI-TOF-MS (matrix assisted laser desorption ionization time-of-flight mass spectrometry) enable high-throughput ultra-trace analysis of polypeptides, enabling identification of trace proteins that could not be detected in the past. It has become a powerful tool in the search for disease-related factors.

  Many bioactive proteins such as peptide hormones, interleukins, and cytokines, which are considered to be pathological biomarkers and pathogenesis-related factors, exist only in trace amounts (<ng / mL). The content ratio is actually nano to pico level as compared with a high content component of high molecular weight such as albumin. In terms of protein size, 70% or more of all types of proteins have a molecular weight of 60 kDa or less, and most of the above-mentioned trace amounts of biomarker proteins are mostly included in this region.

  By the way, when handling proteins, non-specific adsorption of proteins to the resin substrate surface of various analytical instruments used particularly well in the field of biochemistry is always a problem. This non-specific adsorption on the substrate surface not only causes variations in the analysis results due to protein reduction, but also causes serious problems such as loss of the protein to be analyzed, so it is necessary to prevent non-specific adsorption. Generally, the protein reduction rate due to non-specific adsorption depends on the total protein concentration in the solution, and the protein reduction rate increases as the total protein concentration decreases. In particular, when the pathogenesis-related trace component is analyzed by mass spectrometry in the proteome analysis as described above, there is no existing pretreatment apparatus that has been subjected to a process for suppressing nonspecific adsorption. Therefore, the total protein concentration of the fraction obtained by excluding components that inhibit detection is extremely low, and there is a problem of reduction / loss due to nonspecific adsorption of a trace amount of biomarker protein.

  There are two main countermeasures against the problem of loss due to the adhesion of proteins and peptides. One is a method of adding a compound that suppresses adsorption to the biological component solution, and the other is a biological component non-adsorption treatment on the surface of the resin substrate. As a typical method of the former, there is a method of adding a blocking agent. As a blocking agent, a solution of albumin or casein is used, which is a method for suppressing adsorption of useful biological components by competitive adsorption. Because of the competitive adsorption, the concentration of the blocking agent is generally higher than the concentration of useful biological components. Therefore, in analysis applications, there is a risk that the blocking agent inhibits the analysis, and even if a small amount is added, the biological component changes in structure. In addition to blocking, there is a method of adding a surfactant, an inorganic salt, or an organic solvent. However, as with the blocking agent, there is a problem in that the analysis system is inhibited and denaturation due to structural changes of biological components.

  On the other hand, a common non-adsorption treatment on the substrate surface is a hydrophilic treatment on the substrate surface. There are several methods for hydrophilization treatment. For example, Patent Document 1 discloses a method in which a hydrophilic compound such as 2-methacryloyloxyethyl phosphorylcholine copolymer (hereinafter abbreviated as MPC) is introduced into a substrate by coating treatment. Patent Documents 2 to 6 describe methods for introducing a hydrophilic compound by grafting. There is also a method in which hydrophilic functional groups are directly generated on the surface of the substrate, such as reactive ion etching, plasma treatment, and ion cluster beam treatment.

  However, in the conventional base material surface treatment, when the base material subjected to such a treatment comes into contact with a high concentration protein or peptide solution, an effect of suppressing the adsorption of biological components is recognized. When contacted with a solution containing a component, reduction or loss of biological components due to adsorption still occurred, and the level was still not sufficient to solve the problem.

  Furthermore, in the method using a coating treatment with a hydrophilic polymer, when a solvent using a hydrophilic polymer solution is further contacted with the treated substrate, the hydrophilicity may be lowered due to peeling of the coating or the like. Concerned. Further, in a processing apparatus such as analysis or separation, there is a concern that the eluted hydrophilic polymer can be an inhibitor of subsequent analysis.

  Hydrophilization by hydrophilic polymer grafting improves hydrophilicity in proportion to the amount of graft, but if the concentration of the hydrophilic polymer solution to be processed is increased, the hydrophilic polymers are cross-linked three-dimensionally. However, there is a problem that the mobility of the hydrophilic polymer is lowered and the effect of suppressing the adhesion of biological components is lowered. Furthermore, the method described in Patent Document 6 cannot exhibit a sufficient effect under a condition where protein adsorption is likely to occur, such as a low salt concentration.

  In addition, reactive ion etching treatment, plasma treatment, and ion cluster beam treatment can easily hydrophilize the outer surface of the base material or one side of the plate-like base material. It is difficult to make the part to become hydrophilic. Therefore, it is not suitable for making a molded body having a complicated shape hydrophilic by one treatment. Moreover, the adsorption | suction characteristic of the biological component of a base material is dependent on the surface state of the part which contacts a biological component. In general, the higher the hydrophilicity of the surface and the higher the mobility of the hydrophilic molecules immobilized on the surface, the more the biological component is adsorbed onto the substrate surface. It is considered that hydrophilic molecules having high mobility exclude biological components such as proteins and platelets by their molecular motion. Hydrophilization by reactive ion etching treatment, plasma treatment, and ion cluster beam treatment is due to the generation of hydrophilic functional groups such as hydroxyl groups on the substrate surface, that is, by introduction of hydrophilic polymers onto the substrate surface. Compared with hydrophilization, the mobility of hydrophilic molecules is low. Therefore, the effect of suppressing the adhesion of biological components is low and not preferable. Furthermore, since it may become high temperature during a process, since a base material may modify | denature, it is not preferable.

Thus, since the technique of the protein or peptide adsorption | suction suppression process is not established, the resin molded object with few adsorption | suctions is not yet in the world.
JP-T-2002-542163 JP 2003-130882 A JP 58-40323 A Japanese Patent No. 3297707 Japanese Patent Laid-Open No. 61-225653 International Publication No. 06/025352 Pamphlet

  As described above, in the field of manipulating a very small amount of biological material such as clinical proteome analysis, there is a problem that protein is lost due to non-specific adsorption of the protein or peptide to the surface of the molded body, and stable treatment / analysis cannot be performed. In order to perform stable processing and analysis, it is essential to suppress nonspecific adsorption of proteins to the surface of the base material, and polystyrene, polyethylene, polypropylene, polycarbonate, polyvinyl chloride, etc., which are particularly often used in this field It is a problem to provide a technique for suppressing adsorption to the molded resin.

In order to solve the above problems, the present invention employs any of the following means.
(1) The resin molded body includes a step of immersing in an aqueous solution of a nonionic surfactant, and a step of irradiating the resin molded body in a state of being immersed in said aqueous solution, it said in the aqueous surfactant For the treatment of proteins and / or peptides on which a nonionic surfactant is immobilized, wherein the concentration of the agent is in the range of 0.05 to 500 times the critical micelle concentration at 25 ° C. of the surfactant The manufacturing method of the resin molding to use for .
(2) The method for producing a resin molded body according to (1), wherein the aqueous solution contains water-soluble inorganic salts at a concentration of 50 mmol / L to 300 mmol / L.
(3) The manufacturing method of the resin molding as described in said (1) or (2) whose said surfactant is what is represented by Formula 1.

(4) The method for producing a resin molded body according to (3), wherein R ₂ has 2 carbon atoms.
(5) The manufacturing method of the resin molding as described in said (3) or (4) whose number of n is 5-80.
(6) the number of carbon atoms of _{R 1} is 5 to 25, wherein (3) to the production method of the resin molded body according to any one of (5).
(7) The method for producing a resin molded body according to any one of (3) to (6), wherein R ₁ is R ₄ -A-, and R ₄ has 7 to 10 carbon atoms.
(8) The manufacturing method of the resin molding in any one of said (1)-(7) whose HLB of the said surfactant is 10 or more.

  According to the present invention, it is possible to obtain a molded resin that hardly adsorbs biological components, particularly proteins and peptides, on the surface. And according to this resin molding, the loss of the protein etc. made into the objective of analysis can be decreased.

  The resin molding manufactured by this invention is used suitably for the process of protein and / or a peptide. The treatment here means, in particular, an operation for handling proteins and / or peptides performed in the fields of biochemistry, biology, analytical chemistry, agriculture, forestry and fisheries, food, medicine, pharmacy, and the like. In addition to storage, preservation, collection, and dispensing of peptides or solutions containing them, operations such as reaction, analysis, separation, purification, concentration, and drying are also included. Therefore, the shape of the molded body is not particularly limited, and various shapes such as yarns, hollow fibers, fibers, knitted fabrics, films, flat membranes, hollow fiber membranes, particles, tubes, rods, containers, and the like can be used. It is good.

  The type of resin is not particularly limited, and vinyl polymers such as polyethylene, polypropylene, polyvinyl chloride, polyvinylidene chloride, polystyrene, polyvinyl acetate, polyacrylate, polymethacrylate, polymethyl methacrylate, polyacrylonitrile, polyisoprene, polybutadiene or the like Acrylic polymers, polyamide polymers such as nylon, polyimide polymers, polyurethane polymers, polyethylene terephthalate, polybutylene terephthalate, polyester polymers such as polylactic acid, polyglycolic acid, polytetrafluoroethylene, polyvinylidene fluoride, perfluoro Fluoropolymers such as polymers, polycarbonate, polyphenylene oxide, polyphenylene sulfide, polysulfone, polyethylene Terusuruhon, polyether ether ketone, silicone resin, natural rubber, cellulose, is suitably selected from cellulose acetate. Moreover, the copolymer which consists of the said polymer, and resin formed by blending the said polymer may be sufficient.

  In the present invention, the weight average molecular weight, number average molecular weight, molecular weight polydispersity and crystallinity of the resin are not particularly limited, and any resin can be used as long as it can be molded. The molded body may be composed only of a resin, or may be a molded body in which the resin is localized on the surface.

  The resin molded body as described above is used, for example, for protein processing. When protein is dissolved in water, a hydrophilic domain is present on the surface of the protein molecule, and a hydrophobic domain is present inside the protein. Existing. And when protein contacts a hydrophobic base material, an internal hydrophobic domain will be exposed on the surface, and it is thought that it adsorb | sucks to a base material by hydrophobic interaction. Therefore, hydrophilization of the substrate surface is effective for suppressing protein adsorption.

  Therefore, in the present invention, the resin molded body as described above is brought into contact with an aqueous solution of a nonionic surfactant, and the surfactant is physicochemically adsorbed on the surface of the resin molded body. In the present invention, the term “liquid contact” refers to bringing the aqueous solution of the surfactant into contact with the surface of the resin molding. The method is not particularly limited, and includes a method of immersing the resin molded body in the aqueous solution, a method of spraying the aqueous solution onto the molded body, and the like. A dipping method is preferred.

  In the present invention, it is essential that the nonionic surfactant is water-soluble in a temperature range of 1 to 100 ° C. It is sufficient that the surfactant does not cause precipitation in an aqueous solution, and the surfactant may be dissolved as nanospheres or microspheres such as micelles and liposomes.

  Examples of nonionic surfactants include organic compounds represented by Formula 1.

These nonionic surfactants are adsorbed on the surface of the resin molded body through a hydrophobic R ₁ segment by contacting the resin molded body with an aqueous solution, while the surface is treated with hydrophilic polyoxy Hydrophilization with alkylene segment (OR ₂ ) _n . In addition, since the polyoxyalkylene segment (OR ₂ ) _n extends into the protein solution, in addition to the effect of hydrophilization, an excluded volume effect due to micro-Brownian motion of the hydrophilic polymer chain is also exhibited. Therefore, the resin molding obtained according to the present invention can exhibit a protein adsorption suppressing effect.

  The non-adsorbability of proteins and peptides depends on the amount of nonionic surfactant that is physicochemically adsorbed before irradiation and chemically immobilized by irradiation. Therefore, it is important to appropriately control the adsorption amount of the nonionic surfactant in the liquid contact process.

One factor that determines the amount of surfactant adsorbed is HLB (hydrophilic lipophilic balance). The HLB here can be theoretically calculated from the chemical structure by the Griffin method as follows. In addition, the hydrophilic part of the water-soluble compound in the following formula refers to a segment of (OR ₂ ) _n —OR _{3 in} the case of formula (1).

  If the HLB theoretical value is too low, the hydrophilicity is insufficient and the solubility in water is lowered, and if it is too high, the lipophilicity is insufficient and the adsorptivity to the surface of the resin molding is reduced. Therefore, HLB is preferably 10 or more and 20 or less, and most preferably 12 or more and 19 or less.

In addition, the nonionic surfactant represented by Formula 1 determines the solubility in water and the dissolved state in water depending on the balance between the hydrophilic polyoxyalkylene segment and the hydrophobic R ₁ segment. R _{1 has} a chemical structure of a linear or branched alkyl group having 1 to 30 carbon atoms, an alkenyl group, an alkynyl group, or R ₄ -A- (wherein R ₄ is a straight chain having 1 to 18 carbon atoms or The branched alkyl group, alkenyl group or alkynyl group, and A is a phenylene group) are not particularly limited. However, when the number of carbon atoms is small, the interaction with the resin surface becomes weak, and when the number of carbon atoms is large, water solubility at room temperature is achieved. Decreases. Therefore, the carbon number of R ₁ is preferably 5-25. Further, when _{R 1} is _R 4 -A-, in terms of availability, in addition to the above reasons, it is preferable number of carbon atoms of _{R 4} is 7 to 10.

  When the degree of polymerization of the polyoxyalkylene segment is too short, water solubility at room temperature is lowered, and when it is too long, the interaction with the resin surface becomes weak. Among these, the upper limit is preferably 80 or less, and more preferably 50 or less. On the other hand, the lower limit is more preferably 5 or more.

The alkylene chain R ₂ of the polyoxyalkylene segment has 2 or 3 carbon atoms, and the polyoxyalkylene is polyoxyethylene or polyoxypropylene. In particular, polyoxyethylene is preferable because of its high affinity with water.

R ₃ at the end of the polyoxyalkylene chain is an H group, a CH ₃ group or a CH ₂ CH ₃ group, and the H group having the highest hydrophilicity is most preferred.

In the above formula 1, when R ₁ is R ₄ -A-, R ₄ and the polyoxyalkylene segment may be bonded to any position of the phenylene group A, but they are highly stable and easy to synthesize. Most preferred is a para-substituted product at the 1,4-position. In the phenylene group A, the position where R ₁ and the polyoxyalkylene segment are not bonded may be substituted with other substituents as long as the water solubility is not changed.

Specific examples of the nonionic surfactant represented by Formula 1 include Triton X-45, Triton X-100, Triton X-114, Triton X-165, Triton X-200, Triton X-305, Triton X -405, Triton X-705, Triton N-60, Triton N-101, Triton N-111, Triton N-150, Polyoxyethylene (8) Octylphenyl Ether, Polyoxyethylene (9) Octylphenyl Ether, Polyoxyethylene (10) Octylphenyl Ether, Polyoxyethylene (5) Nonylphenyl Ether, Polyoxyethylene (10) Nonylphenyl Ether, Polyoxyethylene (15) Nonylphenyl Ether, Polyoxyethylene (20) Nonylphenyl Ether. Incidentally, these are, there is _{R 1} is _R 4 -A- in Formula 1, the carbon number of _{R 4} is 8 or 9, the number of repetitions n of the polyoxyalkylene segments _{(OR 2) n} is from 10 to 70, The alkylene chain R ₂ in the segment has 2 carbon atoms and R ₃ is H. R ₄ and the polyoxyalkylene segment are bonded to the para-positions 1 and ₄ with respect to the phenylene group.

Specific examples of other nonionic surfactants include Brij30, Brij35, Brij56, Brij58, Brij78, Brij97, Brij98, Polyoxyethylene (6) Decyl Ether, Polyoxyethylene (9) Decyl Ether, Polyoxyethylene (12) Decyl Ether , Polyoxyethylene (20) Cetyl Ether, Polyoxyethylene (10) Dodecyl Ether, Polyoxyethylene (23) Lauryl Ether, Polyoxyethylene (7) Oleyl Ether, Polyoxyethylene (10) Oleyl Ether, Polyoxyethylene (20) Oleyl Ether, Polyoxyethylene (50) Oleyl Ether, Examples include Polyoxyethylene (4) Stearyl Ether and Polyoxyethylene (20) Stearyl Ether. Note that these are the number of carbon atoms of _{R 1} in Formula 1 is 10 to 18, the polyoxyalkylene segments _{(OR 2)} repetition number n in the _n is 6 to 23 carbon atoms in the alkylene chain _{R 2} in the segments 2, R ₃ is H.

  When such a nonionic surfactant is adsorbed on the resin surface, if the concentration in the aqueous solution is too low, the absolute amount sufficient to exert the adsorption-inhibiting effect is insufficient. In addition to easily reducing the reaction efficiency of radicals binding to the surface, the excluded volume effect of the polyoxyalkylene segment is difficult to function effectively. Therefore, the concentration in the aqueous solution is preferably in the range of 0.05 to 500 times the critical micelle concentration of the nonionic surfactant, and most preferably in the range of 0.1 to 200 times. .

The critical micelle concentration here can be evaluated, for example, as follows.
[Measurement condition]
Measuring device: CBVP-A3; manufactured by Kyowa Interface Science Co., Ltd. (or no problem as long as the same result can be obtained under the same conditions)
Test room temperature: 25 ° C
Test room humidity: 60%
Plate: Platinum plate The surface tension was measured under these conditions, and the obtained surface tension was plotted against the concentration (logarithmic concentration) of the water-soluble nonionic surfactant. Determine the concentration (critical micelle concentration).

  Further, for the same reason, the concentration of the water-soluble nonionic surfactant is preferably 0.001% by weight or more and 1% by weight or less, particularly 0.01% by weight or more and 1% by weight or less. Most preferred.

  Furthermore, in the production method of the present invention, since water-soluble nonionic surfactant is bound to the surface of the molded resin using radiation, water-soluble inorganic salts are allowed to coexist in the aqueous solution of the surfactant. preferable. Water-soluble inorganic salts have the effect of enhancing the hydrophobic interaction between the surfactant and the surface of the resin molding. The water-soluble inorganic salts are not particularly limited, and lithium, sodium, potassium, calcium, magnesium, ammonium, iron, zinc hydrochloride, sulfate, carbonate, phosphate and the like are preferably used. The concentration of the water-soluble inorganic salt is not particularly limited, but if it is too low, the effect of strengthening the hydrophobic interaction between the surfactant and the surface of the molded resin will be reduced, and if it is too high, the surfactant will be dissolved. Reduce sex. Therefore, it is preferably 50 mmol / L or more and 300 mmol / L or less, and most preferably 100 mmol / L or more and 300 mmol / L or less with respect to the aqueous solution.

Subsequently, in the present invention, the nonionic surfactant adsorbed on the surface of the resin molded body as described above is chemically bonded to the surface of the resin molded body. Bonding is performed using radiation. That is, the resin molded body and the above-described surfactant are bonded by irradiating the surface of the resin molded body after being in contact with the above-described aqueous solution with radiation. Radiation energy generates active hydroxy radicals in water, and these radicals draw out the resin or the water-soluble nonionic surface active hydrogen to generate new radicals, causing the radical reaction to proceed in a chain and the resin. Bonding occurs on the surface of the compact.
As the radiation, α rays, β rays, γ rays, X rays, ultraviolet rays, electron beams and the like are used. In particular, electromagnetic waves such as γ rays and electron beams have been widely used for sterilization and the like in recent years because of their simplicity. The radiation dose is preferably in the range of 0.01 kGy or more and 100 kGy or less from the viewpoint of the efficiency of bonding to the surface and prevention of deterioration of the resin base material, in the range of 0.1 kGy or more and 50 kGy or less, particularly 0.5 or more and within 40 kGy Most preferably,

  By these steps, the nonionic surfactant is chemically bonded to the surface of the resin molded body by a covalent bond. Therefore, the surface which has the characteristics that a surfactant does not elute and an adsorption | suction suppression effect continues is obtained.

The surfactant bonded to the surface of the resin molding is detected by chemical structure analysis. This utilizes the fact that the surface of the molded body consists only of a resin, whereas the surfactant has a polyoxyalkylene group. For example, ion fragments specific to polyalkylene oxide groups can be detected by TOF-SIMS (time-of-flight secondary ion mass spectrometry). Moreover, when this surfactant has a phenylene group, the signal resulting from the 1100-1300 cm < ^-1 > carbon-oxygen bond of an ATR-IR spectrum is a peculiar signal which is not seen in resin, such as a polypropylene and a polystyrene.

Hereinafter, the present invention will be described in detail with experimental examples, but the scope of the present invention is not limited to these experimental examples. <Method of evaluating adsorption performance using β ₂ -microglobulin>
The protein adsorption evaluation on the substrate surface will be described in the case where an adsorption test is conducted with a solution of human β ₂ -microglobulin (SIGMA sales, Cat. No. M4890) (hereinafter abbreviated as β2-MG).

  Protein solution of 25 mmol / L ammonium bicarbonate aqueous solution (pH 8.2) adjusted to 500 ng / ml of β2-MG and 500 ng / ml of human serum albumin (SIGMA sold, Cat. No. A1653) (hereinafter abbreviated as HSA) (Hereinafter referred to as protein solution A). Since the protein in the protein solution A is also adsorbed in the container used for the preparation, the container used for preparing the protein solution must be prepared in advance with bovine serum albumin (Nacalai Tesque Sales, Cat. No. 01863-35) , Abbreviated as BSA). The container blocking operation was performed after leaving a centrifuge tube (Greiner Bio-One GmbH, CELLSTAR TUBES, 15 mL) in a 1% BSA phosphate buffered saline (hereinafter abbreviated as PBS) solution for 30 minutes. The centrifuge tube was washed 3 times with PBS and 3 times with distilled water. The protein solution A prepared with the centrifuge tube thus blocked was used in the adsorption experiment as follows.

  100 μl of protein solution A was added to a test tube as a resin molding and left at 26 ° C. for 1 hour. One hour later, the protein solution in the test tube was collected, and a 10-fold diluted solution of 1% BSA in PBS was used for measurement of β2-MG concentration (c). The β2-MG concentration (b) was also measured for the protein solution A before being dispensed into the resin test tube.

The β2-MG concentration (b) was measured with a β2-MG measurement kit (Wako Pure Chemical Industries, Ltd., Glazyme β ₂ -microgloblin EIA TEST, Code. 305-11011) according to the manual attached to the kit. A part of the attached manual was modified and a reaction vessel previously blocked with 1% BSA in PBS for 30 minutes was used for the first reaction. The protein adsorption rate (a) was calculated by the following equation, and the case where the adsorption rate was 50% or less was defined as a non-adsorbed surface.

<Measurement method of critical micelle concentration>
[Measurement condition]
Measuring device: CBVP-A3 (manufactured by Kyowa Interface Science Co., Ltd.)
Test room temperature: 25 ° C
Test room humidity: 60%
Plate: Platinum plate An aqueous solution of a nonionic surfactant was put in a container, and the surface tension was measured according to the attached manual of CBVP-A3. For various concentrations of nonionic surfactant, the surface tension is measured in the same manner, and the value of each surface tension is plotted against the concentration of nonionic surfactant (logarithmic concentration). The critical micelle concentration of the nonionic surfactant was determined by determining the lowest concentration.

(Experimental example 1)
Resin test tube ("5 ml Polypropylene Round-Bottom Tube" manufactured by BECTON DICKINSON) is added to 100 ml of 0.001, 0.01, 0.1 and 1% Triton X-100 (Wako Pure Chemicals, Cat No.168-11805) aqueous solution. Five of each were immersed and irradiated with γ rays. The absorbed dose of gamma rays was 25 kGy. The test tube was removed from the aqueous solution, washed 3 times with 500 ml of running water and air dried at room temperature. Of these test tubes, three of each concentration was subjected to the human β2-MG adsorption test, and the average value of the adsorption rate was determined. The conditions are shown in Table 1, and the results are shown in Table 2. (Experimental example 2)
A test tube made of resin ("5ml Polypropylene Round-Bottom Tube" manufactured by BECTON DICKINSON) is used in polyoxyethylene (10) nonylphenyl ether (Wako Pure Chemicals Cat No.320-33722) at concentrations of 0.0001, 0.001, 0.001, 0.01 and 0.1%. Five of each were immersed in 100 ml of an aqueous solution, treated in the same manner as in Experimental Example 1, and subjected to a human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 3)
Each resin test tube (“5 ml Polystyrene Round-Bottom Tube” manufactured by BECTON DICKINSON) is added to 100 ml of 0.001, 0.01, 0.1, and 1% Triton X-100 (Wako Pure Chemicals Sales Cat No.168-11805) aqueous solution. Five pieces were immersed in each, treated in the same manner as in Experimental Example 1, and subjected to a human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 4)
A resin test tube ("5 ml Polystyrene Round-Bottom Tube" manufactured by BECTON DICKINSON) is added to 100 ml of 0.001, 0.01, 0.1, 1, 2 and 5% TritonX-705 (SIGMA sales Cat No. X70570-100ML) aqueous solution. Five of each were immersed, treated in the same manner as in Experimental Example 1, and subjected to a human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 5)
Resin test tubes ("5 ml Polystyrene Round-Bottom Tube" made by BECTON DICKINSON) are added to 100 ml of 0.0001, 0.001, 0.01, 0.1 and 1% TritonX-405 (SIGMA sales Cat No. X405-100ML) aqueous solution each 5 The samples were immersed one by one, treated in the same manner as in Experimental Example 1, and subjected to a human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 6)
Resin test tube (BECTON DICKINSON's “5ml Polystyrene Round-Bottom Tube”) in 100 ml of 0.01, 0.1, 1, 2, and 5% Triton X-45 (SIGMA Cat No. X45-100ML) aqueous solution Five of each were immersed, treated in the same manner as in Experimental Example 1, and subjected to a human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 7)
Five test tubes made of resin (BECTON DICKINSON's “5 ml Polystyrene Round-Bottom Tube”) in 100 ml of 0.0001, 0.001, 0.01, 0.1 and 1% Brij 58 (SIGMA sales Cat No. P5884-100G) aqueous solution each. The samples were immersed one by one, treated in the same manner as in Experimental Example 1, and subjected to human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 8)
5 each of resin test tubes (5 ml Polypropylene Round-Bottom Tube made by BECTON DICKINSON) in 100 ml of 0.0001, 0.001, 0.01, and 0.1% concentrations of Brij 58 (SIGMA sales Cat No. P5884-100G) The sample was immersed, treated in the same manner as in Experimental Example 1, and subjected to a human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 9)
A test tube made of resin ("5 ml Polystyrene Round-Bottom Tube" manufactured by BECTON DICKINSON) in an aqueous solution of 0.1% Triton X-100 (Wako Pure Chemicals Cat No.168-11805) containing 100, 200 and 300 mmol / L sodium chloride 5 pieces each were immersed in 100 ml, treated in the same manner as in Experimental Example 1, and subjected to human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 10)
Resin test tube ("5 ml Polystyrene Round-Bottom Tube" manufactured by BECTON DICKINSON) is added to 100 ml of 0.1% Triton X-305 (SIGMA sales Cat No. X305-500ML) aqueous solution containing 100, 200 and 300 mmol / L sodium chloride. Each was immersed in five pieces, treated in the same manner as in Experimental Example 1, and subjected to a human β2-MG adsorption test. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 11)
Since an attempt was made to prepare an aqueous solution of Triton X-15 (SIGMA sales Cat No. X15-500ML) but it did not dissolve, the same γ-ray treatment of the resin test tube as in Experimental Example 1 could not be performed.

(Experimental example 12)
Five resin test tubes (“5 ml Polypropylene Round-Bottom Tube” manufactured by BECTON DICKINSON) were immersed in 100 ml of 0.01% Triton X-100 aqueous solution. Without γ-irradiation, the test tube was treated in the same way as the post-treatment after γ-irradiation in Experimental Example 1. Three of these test tubes were subjected to the human β2-MG adsorption test, and the average value of the adsorption rate was determined. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 13)
Five resin test tubes (“5 ml Polystyrene Round-Bottom Tube” manufactured by BECTON DICKINSON) were immersed in 100 ml of 0.01% Triton X-100 aqueous solution. Without γ-irradiation, the test tube was treated in the same way as the post-treatment after γ-irradiation in Experimental Example 1. Three of these test tubes were subjected to the human β2-MG adsorption test, and the average value of the adsorption rate was determined. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 14)
According to the method described in Patent Document 6, 5 test tubes made of resin ("5 ml Polyprpylene Round-Bottom Tube" manufactured by BECTON DICKINSON) are immersed in 100 ml of 0.1% aqueous solution of polyvinyl alcohol (Poval 205, manufactured by Kuraray) without critical micelle concentration. And irradiated with γ rays. The absorbed dose of gamma rays was 25 kGy. The resin test tube was taken out of the polyvinyl alcohol aqueous solution, washed with 500 ml of running water, and dried in an oven at 70 ° C. for 1 hour. Three of these test tubes were subjected to the human β2-MG adsorption test, and the average value of the adsorption rate was determined. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 15)
According to the method described in Patent Document 6, 5 test tubes made of resin (“5 ml Polystyrene Round-Bottom Tube” manufactured by BECTON DICKINSON) are immersed in 100 ml of 0.1% aqueous solution of polyvinyl alcohol (Poval 205, manufactured by Kuraray) without critical micelle concentration. The absorbed dose of γ rays was 25 kGy.The resin test tube was taken out of the polyvinyl alcohol aqueous solution, washed with 500 ml of running water, and dried in an oven at 70 ° C. for 1 hour. The book was subjected to a human β2-MG adsorption test, and the average value of the adsorption rate was determined, the conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 16)
Five resin test tubes ("5 ml Polypropylene Round-Bottom Tube" manufactured by BECTON DICKINSON) were washed with 500 ml of running water and air-dried at room temperature. Three of these test tubes were subjected to the human β2-MG adsorption test, and the average value of the adsorption rate was determined. The conditions are shown in Table 1, and the results are shown in Table 2.

(Experimental example 17)
Five resin test tubes (“5 ml Polystyrene Round-Bottom Tube” manufactured by BECTON DICKINSON) were washed with 500 ml of running water and air-dried at room temperature. Three of these test tubes were subjected to the human β2-MG adsorption test, and the average value of the adsorption rate was determined. The results are shown in Table 1.

  As is apparent from Tables 1 and 2, as a result of the human β2-MG adsorption test, the amount of human β2-MG adsorbed is smaller in the case of the present invention than in the comparative example, and the adsorption of trace biological components is suppressed and the recovery rate is high. It is effective.

  The production method of the present invention is very useful in terms of preventing adsorption loss when processing and analyzing trace amounts of proteins and / or peptides, and is particularly useful for medical, particularly human diseases, when used for proteome analysis. Contribute to the discovery of

Claims (8)

The resin molded body includes a step of immersing in an aqueous solution of a nonionic surfactant, and a step of irradiating the resin molded body in a state of being immersed in the aqueous solution, the concentration of the surfactant in the aqueous solution Is a resin used for the treatment of proteins and / or peptides on which a nonionic surfactant is immobilized, which is in the range of 0.05 to 500 times the critical micelle concentration at 25 ° C. of the surfactant A method for producing a molded body. The method for producing a resin molded body according to claim 1, wherein the aqueous solution contains water-soluble inorganic salts at a concentration of 50 mmol / L to 300 mmol / L. The manufacturing method of the resin molding of Claim 1 or 2 whose said surfactant is what is represented by Formula 1.

The number of carbon atoms in R ₂ is 2, the manufacturing method of the resin molded body according to claim 3. The manufacturing method of the resin molding of Claim 3 or 4 whose number of n is 5-80. The number of carbon atoms of R ₁ is 5 to 25, a manufacturing method of the resin molded body according to any one of claims 3-5. R ₁ is _R 4 -A-, the carbon number of _{R 4} is 7 to 10, a manufacturing method of the resin molded body according to any one of claims 3-6. The manufacturing method of the resin molding in any one of Claims 1-7 whose HLB of the said surfactant is 10 or more.